Detection of doping control sample substitutions via single nucleotide polymorphism-based ID typing.

Area of Science:

• Forensic Science
• Genetics
• Biochemistry

Background:

• Sample authenticity is crucial in sports drug testing to prevent doping violations.
• Sample swapping, where an athlete submits another person's sample, is a known method of cheating.
• Existing methods need robust tools to detect such sample manipulations.

Purpose of the Study:

• To assess the utility of a single nucleotide polymorphism (SNP)-based sample authentication method.
• To evaluate a multi-target SNP panel for detecting doping control sample manipulation.
• To determine the effectiveness of this method for urine and dried blood spot samples.

Main Methods:

• Development and application of a multi-target SNP panel with 44 SNPs, gender, and quality control markers.
• Utilized multiplex polymerase chain reaction and single base extension reactions.
• Analyzed DNA profiles using MALDI-TOF MS for sample authentication.

Main Results:

• The SNP panel reliably typed and matched urine and dried blood spot samples to reference whole blood samples.
• Efficient typing of urine samples was dependent on sample quality and quantity, with robust profiling achieved at 12 μl.
• The assay demonstrated a high-throughput capability with a turnaround time of approximately 11 hours.

Conclusions:

• SNP-based DNA typing using MALDI-TOF MS is a viable, high-throughput method for doping control sample authentication.
• This SNP profiling can complement existing steroid profile analysis to confirm sample manipulations.
• The method supports quality control processes in high-throughput anti-doping laboratories.